|
| Status |
Public on Sep 30, 2021 |
| Title |
∆rrpAB rep4 |
| Sample type |
SRA |
| |
|
| Source name |
not applicable
|
| Organism |
Campylobacter jejuni |
| Characteristics |
strain: 11168H
genotype: rrpAB double mutant
culture method: cell suspension culture (6 hrs growth)
|
| Growth protocol |
RNA-Seq was used to identify differentially expressed genes between wild type strains, ∆rrpA and ∆rrpB mutants at mid-log (~6 hrs) of growth. 11168H wild type, 11168H∆rrpA, 11168H∆rrpB and 11168H∆rrpAB were plated out on BA plates and incubated at 37°C under microaerobic conditions for 6 hrs. 11168H is hypermotile variant of strain 11168 which is the background strain for the samples used in the study.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted from the pelleted bacteria cells using PureLink™ RNA Mini Kit (Invitrogen), following manufactures protocol. RNA was isolated from five independent experiments performed on different days Ribosomal RNA was depleted using Ribominus (Invitrogen)
RNA libraries were prepared for sequencing using standard Illumina protocols
|
| |
|
| Library strategy |
RNA-Seq |
| Library source |
transcriptomic |
| Library selection |
cDNA |
| Instrument model |
Illumina MiSeq |
| |
|
| Description |
11168H_rrpAB_biological_4
|
| Data processing |
Raw reads were obtained from an Illumina Miseq paired-end sequencing platform (Illumina).
The paired-end reads were trimmed and filtered using Sickle v1.200 by using a sliding window approach and trimming the reads where the average base quality drops below 20. Only the reads that were above 10 bp length were kept after trimming. Bowtie2 was used to map the reads against the reference sequences; C. jejuni strains 11168H assembly GCA_900117385.1. This generated the mapping in SAM format which were converted to BAM format using Samtools and were index sorted. We used gffread from cufflinks suite to convert annotations from GFF to GTF format. These were used in Bedtools (with multicov-bams option)with the mapped reads to generate transcript counts per samples. A shell utility by the authorswas then used to collate all these transcripts into a n=20 samples x p=1,628 transcripts for 11168H strain. Statistical analysis was performed in R using the combined data generated from the bioinformatics as well as meta data associated with the study (multifactorial design). We used DESeq DataSet from Matrix()function fromDESeq2 package with the adjusted p-value significance cut-off of 0.05 and log fold change cut-off of 1.5. This function uses negative binomial GLM to obtain maximum likelihood estimates for the transcripts log fold change between the two conditions.
Genome_build: C. jejuni strains 11168H assembly GCA_900117385.1.
Supplementary_files_format_and_content: Excel file include RPKM values for each Sample
Supplementary_files_format_and_content: Matrix table with raw gene counts for every gene and every sample
|
| |
|
| Submission date |
May 12, 2021 |
| Last update date |
Sep 30, 2021 |
| Contact name |
Fauzy Nasher |
| E-mail(s) |
fauzy.nasher1@lshtm.ac.uk
|
| Organization name |
London School of Hygiene and Tropical Medicine
|
| Street address |
Keppel Street,
|
| City |
LONDON |
| ZIP/Postal code |
WC1E 7HT |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL22824 |
| Series (1) |
| GSE174333 |
MdaB and NfrA, two novel reductases important in the survival and persistence of the major enteropathogen Campylobacter jejuni |
|
| Relations |
| BioSample |
SAMN19117158 |
| SRA |
SRX10856732 |
